PROJECT SUMMARY/ABSTRACT Genomics and Bioinformatics Core (Core 2) The overall objective of the Genomics and Bioinformatics Core is to provide PPG investigators access to state- of-the-art genomics and bioinformatics technologies via both performance and teaching-based approaches. The Genomics and Bioinformatics Core works with PPG investigators to help design, execute, and interpret experiments that maximize significance of the data obtained while consuming the minimal resources required. The Genomics and Bioinformatics Core is directed by Drs. Patrick Gallagher, PI, and Dr. Vincent Schulz, Scientific Director. The Core performs a variety of assays that leverage the power of ultrahigh throughput sequencing. RNA expression-based techniques include RNA-seq and single cell transcriptome profiling. Genome wide analysis of DNA methylation status is assessed via enhanced reduced representation bisulfite sequencing and of chromatin accessibility via ATAC sequencing. Histone architecture is assessed via ChIP-seq. Control of gene expression at the level of translation is assessed via ribosomal profiling. A critical aspect of these studies is application of strategies to perform analyses on small numbers of cells, allowing protocol development for study of unamplified, primary human erythroid cells. Comprehensive bioinformatic services provided include analyses of RNA expression, DNA methylation, chromatin accessibility, and histone architecture. The Core also provides analysis of protein expression via ribosome profiling and via analysis of proteomic data sets. The influence of genetic variation is characterized in transcriptome, epigenetic, and proteomic data sets. Various strategies are utilized to integrate all of these data types to gain a more complete understanding of erythropoiesis. For these studies, comprehensive quality control procedures are in place. An additional aim of the Bioinformatics Core is to provide training and education to PPG investigators and guidance in the use of these technologies, to provide specialized services, and to evaluate and optimize the application of expression, genomic, and proteomic analyses to solve challenging problems confronting hematology researchers. Finally, an ongoing function of the Core is to monitor and adopt, where indicated, the newest and most promising novel bioinformatics programs and algorithms, as well as updates to currently existing programs, for use by PPG investigators.